Method of implanting heavy metal such as nobel metal, e.g.gold, and metal for use in implantation

ABSTRACT

The present invention relates to a method and a heavy metal, such as noble metal. E.g. gold, for implantation in vivo or in vitro in local areas in humans or animals, the gold is provided with a large surface to ensure ample release of gold ions. The heavy metal e.g. gold may be admixed to solids, (tablets, powders) liquids (solutions, lung sprays, eye drops, nose spray) or cremes, depending on how it is going to be used, on the different external or internal epithelial surfaces. Form implantation in tissues including the brain, the gold is provided as pieces of gold having a mass and surface adapted to the purpose, where the surface of the piece of gold is larger than a cylindrical piece of gold having the same mass. In a preferred embodiment, the pieces of gold are wound as springs.

[0001] The heavy metals such as noble metals gold and silver and mercuryhave been used since ancient times as cures for a wide range of diseasesincluding tuberculosis, but only within the last 50 years, a morerational basis for the therapeutic use of gold and silver has emerged.

[0002] However, the use of metallic gold implants as a paramedicalremedy for the treatment of rheumatism and pain is still consideredirrational, because a scientific explanation of the postulatedbeneficial effects is lacking.

[0003] The authorized and scientifically well-founded use of gold in themedical world is mainly performed with various gold-thio compounds suchas gold sodium thiomalate and recently Auronofin R, a drug suited fororal treatment of rheumatoid arthritis.

[0004] Already quite early on it was suggested that gold inhibits thelysosomal enzymes of phagocytotic cells in the inflamed synovial tissue.

[0005] Although knowledge of the effect of gold ions on the cellularimmuno response is still sparse, it is recognized that gold ions arepowerful inhibitors of macrophages and polymorphonuclear leucocytes, andthe ability of the above-mentioned gold compounds to suppressinflammation in rheumatic joints was established a long time ago.

[0006] Gold ions are believed to inhibit antigen processing and tosuppress NF-kappa B binding activity and I-kappa B-kinase activation,resulting in a reduced production of proinflammatory cytokines.

[0007] The use of gold implants originates from acupuncture where goldneedles seem to have been used quite often.

[0008] In the early 1970s some US veterinarians started to treat dogssuffering from hip dysplasia with gold implants, and since then severalveterinary surgeons and medical doctors have implemented the techniqueon their patients.

[0009] The possibility of silver-enhancing minuscule metallic goldclusters in tissues by autometallographic (AMG) silver enhancement wasrealized in 1981 and was soon implemented as a tool for enhancingcolloidal gold particles bound to antibodies and enzymes.

[0010] Gold ions bound in gold complexes will give rise to nano sizedclusters of metallic gold if subjected to reduction e.g. by radiatingtissue with UV light.

[0011] It is realised that gold implants release gold ions, most likelyby macrophage caused oxidation of the surface, which bind to speciessuch as cyanide or sulfhydryl groups in tissue adjacent to the implant.The gold-loaded molecules are then phagocytosed by cells close to thegold implants and finally accumulated in the lysosomes.

[0012] After reduction of the tissue sections, the resultingnanoclusters of gold atoms are visualized by AMG.

[0013] It is therefore hypothesized that the surface of the goldimplants stimulates a reaction from the immune system, causing anoxidative liberation of gold ions that bind to the above-mentionedmolecules.

[0014] The released gold ions, possibly AU(I), may exert theirinhibitory effect on inflammation either directly by harming peptides orproteins by reacting chemically with them, or they may have to beconverted to aurocyanide ions first. If in excess, Au(I) or Au(CN)⁻ ₂might drift further into the intercellular space, bind to e.g.sulphydryl species and be phagocytosed by mast cells, macrophages andother cells farther away.

[0015] Based on the present knowledge it can be stated that the largerthe surface of the gold implants is, the more gold ions are released andthe farther away cell loaded with gold can be found.

[0016] Since it has thus been shown that it is gold ions released frompure gold that give the desired effect, it is accordingly an object ofthe invention to provide a heavy metal in which the greatest possibleamount of metal ions may be released after implantation. It is likewisean object to apply miniscule gold particles, such as colloidal goldparticles to internal and external surfaces, subjected to immunoreactivereactions with the purpose of damping the immunreaction.

[0017] The object of the invention is achieved by a heavy metal of thetype defined in the introductory portion of claim 1, which ischaracterized in that the metal is formed by one or more pieces e.g. ofgold, having a surface of a piece of metal having a given mass that islarger than the surface of a spherical or cylindrical piece of metalhaving the same mass.

[0018] Hereby, the desired and optimised levels of metal ions areobtained locally in human or animals.

[0019] When, as stated in claim 2, the metal is provided as colloidalgold particles, or the metal is mixed into a solution, or the metal iscreated as carrier substances to be dispensed as drops, or as air soles(sprays) or as ointments or cremes or tablets or capsules, it is ensuredthat the release of gold particles can be adapted to certain treatments,such as internal or external treatment, eye treatments or the like.

[0020] Further advantages in treatments in connection with the examplesmentioned in claim 2 are:

[0021] For treatment of respiratory immunoreactive maladies, it is anadvantage if the colloidal gold containing solution is applied as aspray with a spray device, and for treatment of allergic reactions inthe nose a nose spray can be used.

[0022] For downgrading immunoresponses in the gastrointestinal canal,including the mouth, colloidal gold particles are mixed into the contentof tablets mixtures.

[0023] Finally, for reducing the intensity of allergic skininflammations and eczemas the colloidal gold particles can be dispensedin cremes, ointments or the like.

[0024] A practical provision of gold with a large surface may beachieved as stated in claim 3 in that the threads of gold are wound asspring like coils with one or more windings.

[0025] Likewise, a large surface of the gold may be ensured in that, asstated in claim 4 that the gold implants are created by reticulated orfoiled gold or by foamed gold and as stated in claim 5, that the goldimplants are created of sintered metal particles e.g. gold particles.

[0026] The invention will be illustrated more fully below with referenceto the drawing, in which

[0027]FIG. 1, pictures A-I show micrographs of autometallographically(AMG) developed tissue sections, from tissue samples taken near a goldimplant, while

[0028]FIG. 2, pictures A-D show electron micrographs from gold-implantedrat brain.

[0029] In FIG. 1, pictures A-H show a 5-μm methacrylate embeddedconnective tissue section.

[0030] In FIG. 1, picture I shows an Erpon embedded 3-μm thick brainsection. All sections were counterstained with toluidine blue.

[0031] In picture A, it will be seen that part of a gold grid 1dominates the picture. In the closed mesh windows cells are loaded withsilver-enhanced gold clusters 2.

[0032] In the open window far less cells are stained, see numeral 3.

[0033] In picture B, part of a gold grid mesh is seen at 4 in the upperpart of the picture. The dusty appearance of the tissue close to thegold implant represents gold clusters located outside cells. The twoloaded cells, cf. 5 further away, are believed to be macrophages.

[0034] Picture C shows a rather intense AMG staining of macrophages,mast cells and fibroblasts in the connective tissue from subcutis.

[0035] The gold containing cells, cf. the numeral 6, was located only afew mm from a gold implant.

[0036] Picture D was taken from an AMG stained section of ademineralised piece of the skull bone. The gold implant was placed in adrilled hole in the skull where it became gradually embedded in thebone.

[0037] Newly created bone lamella, cf. numeral 7, encircled theconnective tissue implant connective tissue.

[0038] Macrophages and fibroblasts loaded with AMG grains are visible inthe connective tissue.

[0039] Picture E shows a greater magnification of the square than infig. D, and picture F a greater magnification of the square in C.

[0040] Picture G shows a photomicrograph of mast cells from goldimplanted subcutis.

[0041] It is noted that only part of the mast cell granules containedgold that is indicated by numeral 8.

[0042] Picture H shows a typical fibroblast from connective tissuesurrounding a gold implant for six weeks.

[0043] Picture I is taken from a rat neocortex. The implant was locatedapproximately 1 mm, cf. numeral 10, from the AMG stained astrocyte andneurons.

[0044]FIG. 2, picture A shows two minor interneurons from neocortex.

[0045] Note the exclusive localization of gold in lysome-likeorganelles, cf. numeral 12 in FIG. 2, picture A to C.

[0046] The gold grid based implant was located approximately 1.5 mm fromthe stained neurons.

[0047] Picture B shows gold-loaded neuron from caudate putamen.

[0048] Picture C shows astrocytes adjacent to a capillary (c).

[0049] Picture D shows astrocytes loaded with gold. The AMG grains seemto be free in the cytoplasm, and the nucleus reveals tiny AMG particlesin the euchromatine, cf. the arrows having numeral 14.

[0050] In the following an experiment carried out by the inventor willbe explained:

[0051] Twenty-seven adult rats had threads of pure gold implanted in thebody. The compact threads were 1.25 mm long and 0.5 mm thick, andweighed approximately 5.5 mg. They were cut from a twenty-four caratgold thread. The gold threads were implanted close to arthritic jointsin an attempt to decrease pain and inflammation.

[0052] Another ten rats had implanted coiled-up gold grids with adiameter of 3.0 mm and weighing 0.5 mg. The animals were anaesthetizedwith Nembutal, and the gold threads were implanted by a needle and astiletto into different parts of the body including the brain close tojoints.

[0053] At different intervals of survival (10 days to 3 months) theanimals were reanaesthetized and transcardially perfused with 3%glutaraldehyde in a 0.1 M phosphate buffer.

[0054] The tissues holding the gold threads or grids were excised andplaced in the fixative for at least two hours.

[0055] The tissue blocks were then either frozen on a stage and cut into20-μm sections in a cryostat, or they were cut into 100-μm sections in avibratome.

[0056] Every second glass slide was radiated for 30 min with UV light(wavelength 365 nm) upon which all specimens were AMG developed floatingin the AMG developer. From the developed vibratome sections, areas ofinterest were cut out with razor blade and treated with 0.5% osmiumtetraoxide for 30 minutes, rinsed, dehydrated and embedded in Epon. Someof the tissue blocks were not fixed in osmium, but directly embedded inmethacrylate.

[0057] Every second glass slide with cryostat or methacrylate sectionswas radiated with UV light for 30 min. and placed in a jar, covered bythe AMG developer and placed in a 26° C. water bath.

[0058] A light-tight hood covered the set-up through the 60 minutes ofdevelopment. The AMG developer was then replaced by a 5% thiosulphatesolution for 10 min in order to remove all silver ions from thesections, and ultimately the sections were rinsed several times indistilled water and counterstained with toluidine blue (15).

[0059] Semi-thin sections were cut from the Epon blocks and placed onglass slides. They were counterstained with toluidine blue (1%) andanalyzed in the light microscope. When photos had been taken, selectedsections were reembedded with a drop of Epon on top of a blank Epon.Ultra-thin sections were cut, stained with uranyl and lead, and analysedin an electron microscope.

[0060] Controls included sections from tissues not adjacent to goldimplants and sections treated with a 10% potassium cyanide solution for30 min. Blocks of the tissue surrounding the implants were analyzed withProton Induced X-ray Emission spectroscopy (PIXE). The 20-μm thickcryostat sections were placed on framed micropore polystyrene membranesso thin that the energy loss due to small angle scattering wasnegligible.

[0061] This experiment shows that connective tissue blocks from skin andjoints contained a rim of gold ion-imbibed tissue around the goldimplants, cf. also FIG. 1, pictures A and B. In this juxta-positionedimplant tissue, macrophages and mast cell were the first to showaccumulation of gold, cf. FIG. 1, pictures B-H. already after 14 daysthe first traces of AMG enhanced gold clusters could be observed, andafter one to two months an increasing amount of loaded cells includingfibroblasts were seen. Not all macrophages or mast cells showed anuptake of gold, quite the reverse. Even close to the gold implants somemacrophages and mast cells completely void of gold were observed. Themost heavily loaded cells, whether macrophages or mast cells, showedsigns of degeneration. In cells showing degeneration, EM magnificationsshowed gold accumulations that were not enclosed by a membrane, butseemed to be located free in the cytoplasm. The pattern gave theimpression that lysosomes/vesicles take up increasing amounts of golduntil the enclosing membrane bursts. Cells with free gold accumulationssometimes had gold accumulations in the nucleus. Cells closer to thegold implants were more loaded than cells farther away, and macrophagesand mast cells stained more heavily than fibroblasts. Fibroblasts becameloaded only after long periods of time, approximately two months in thecase of rod-shaped gold threads, but faster if coiled-up gold grids wereused as implants.

[0062] Also the longer the implant had been in the tissue, the morestained cells and the broader the rim of gold-imbibed tissue.

[0063] The coiled-up grid implants, having a substantially increasedsurface compared to the rod-shaped gold threads, released far more goldions, recorded as a drastic increase in the amount of stained cells andin the staining intensity in the individual cells. Kidneys and liversfrom implanted animals were examined in order to test whether gold ionsspread systemically.

[0064] In cases where more than one coiled-up grid was implanted, afaint staining in the proximal tubule of the kidney was recorded in someanimals, while the liver was void of AMG grains.

[0065] If coiled-up grids were implanted directly in the kidney, somerelease of gold ions could be detected in nearby cells, but of all theimplanted tissues those in the kidney caused the least release of goldions.

[0066] The compact gold threads implanted in the brain revealed a tinyrim of imbibed cells closely associated to the implant, but goldion-loaded neurons and glia cells could be traced further from the rim,cf. FIG. 1, picture 1. The coiled-up grid implants resulted in an up toseveral millimetre wide zone of gold-loaded neurons and glia cellsaround the implant. Again some neurons and glia cells were completelyvoid of gold. In a few animals the implant had been embedded inconnective tissue, and no AMG grains were detected in the surroundingneurons and glia cells.

[0067] In both glia cells and neurons gold was found to be located inlysosome-like vesicles, cf. FIG. 2, pictures A-D. The loaded vesicleswere spread throughout the neuronal somata and into the major dendrites.Most of the gold-containing glia cells seemed to be protoplasmaticastrocytes, cf. FIG. 2, pictures C and D, but the possibility thatoligodendroglia and microglia take up gold cannot be excluded at thisearly stage.

[0068] In addition to the fact that AMG staining was observed only intissues relatively close to the gold implants, several controls wereintroduced in order to ensure that the staining was caused by goldclusters: the tissue sections were treated with 10% aqueous potassiumcyanide for 30 minutes. This treatment resulted in a complete removal ofthe catalyst, and the following AMG development resulted in blanksections. The multi-element analysis of cryostat sections with PIXEshowed that the tissue surrounding gold implants contained 38 p.p.m.gold, while tissue taken 1 cm from gold implant was void of gold (<0.39p.p.m).

[0069] Even that the invention has been explained in connection withcertain embodiments, a few remarks concerning possible furtherembodiments are described below:

[0070] 1) The possibility of surface treatment of metal implants e.g.gold and colloidal gold particles with molecules that enhanceldecreasethe amount of released metal ions over time.

[0071] 2) Adhering molecules to such surfaces that support thetherapeutic effects of metal ion release.

[0072] 3) Use of implants or colloidal metallic surfaces as carrier oftherapeutic molecules and ions e.g. molecules that is slowly releasedfrom such surfaces placed on epithelial surfaces or in tissues.

[0073] 4) Use of metallic molecules that ensure that the colloidal metalparticles are taken up by certain types of cells.

[0074] 5) Use of molecules that ensure that the colloidal gold isconcentrated in a particular cell organelle.

[0075] 6) Use of alloys for implantation in humans or animals.

1. A heavy metal, such as noble metal e.g. gold for use in implantationin humans or animals, characterized in that the metal is formed by oneor more pieces e.g. of gold, having a surface of a piece of metal havinga given mass that is larger than the surface of a spherical orcylindrical piece of metal having the same mass.
 2. A heavy metalaccording to claim 1, characterized in that the metal is provided ascolloidal gold particles, or the metal is mixed into a solution, or themetal is created as carrier substances to be dispensed as drops, or asair soles (sprays) or as ointments or crèmes or tablets or capsules. 3.A heavy metal according to claims 1-2, characterized in that the threadsof gold are wound as spring like coils with one or more windings.
 4. Aheavy metal according to claims 1-3, characterized in that the goldimplants are created by reticulated or foiled gold or by foamed gold. 5.A heavy metal according to claims 1-4, characterized in that the goldimplants are created of sintered metal particles, e.g. gold particles.